Table 2. Prediction of residue burial and active-site proximity from Ala and Asp scanning mutagenesis.
Ala*
|
Asp*
|
||||||
---|---|---|---|---|---|---|---|
Class | 0% | 0.1% | 0% | 0.1% | Burial prediction | No. of positions | Accuracy, % |
1 | A | A | A | A | Exposed,† not active site | 40 | 90 |
2 | A | A | I | A | Partially exposed,‡ not active site | 5 | 80 |
3 | A | A | I | I | Buried, not active site or exposed, active site§ | 7 | 86 |
4 | I | A | I | A | Partially exposed‡ | 1 | 100 |
5 | I | A | I | I | Buried, not active site | 9 | 77 |
6 | I | I | I | I | Active site proximal | 4 | 100 |
0% and 0.1% refer to phenotypes of Ala or Asp mutants at 0% and 0.1% arabinose, respectively. A and I refer to active and inactive phenotype, respectively
Represents exposed residues with an ACC > 5%
Represents partially exposed residues with an ACC of 5–15%. Exposed residues can be subdivided into partially (ACC 5–15%) and completely (ACC > 15%) exposed categories
For buried positions, the corresponding Asp mutant will show substantial reductions in protein expression and/or solubility. If the residue is an exposed hydrophobic that is part of the active site, expression and solubility will not be affected